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Related Experiment Videos

Monoclonal autoantibodies recognizing histone variants.

F J van Hemert1, A P van Dam, L J Jonk

  • 1Hubrecht Laboratory, Netherlands Institute for Developmental Biology, Utrecht.

Immunological Investigations
|May 1, 1988
PubMed
Summary

Researchers aimed to create monoclonal antibodies against Ro(SS-A) but unexpectedly developed hybridomas targeting histone H1 and H2B. These antibodies recognize specific histone variants, suggesting they originate from natural autoantibodies rather than immunization.

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Area of Science:

  • Immunology
  • Molecular Biology
  • Biochemistry

Background:

  • Monoclonal antibodies are crucial tools in biomedical research.
  • Developing specific antibodies often involves immunization protocols.
  • Autoantibodies can sometimes be inadvertently generated during antibody production.

Purpose of the Study:

  • To prepare monoclonal antibodies against human Ro/SS-A antigen.
  • To investigate the unexpected reactivity of generated hybridomas.
  • To characterize the specificity of novel anti-histone antibodies.

Main Methods:

  • Immunization of Balb/c mice with purified Ro(SS-A).
  • Fusion of mouse myeloma cells with spleen cells to create hybridomas.
  • Immunoblotting and two-dimensional gel electrophoresis to analyze histone variant specificity.

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Main Results:

  • Monoclonal antibodies against Ro/SS-A were not obtained.
  • Two IgM-producing hybridomas reactive with histone H1 were generated.
  • One hybridoma showed specificity for histone H2B.
  • One anti-histone H1 monoclonal (CLB-ANA 105) recognized H1(0) in mouse and human, and H5/H1 in Xenopus.
  • Another anti-histone H1 monoclonal (CLB-ANA 108) exclusively reacted with mouse H1c.

Conclusions:

  • The generated monoclonal antibodies were not against Ro/SS-A but against histone variants.
  • The specificities suggest these antibodies are derived from naturally occurring autoantibodies, not the immunization process.
  • These findings offer insights into the origins of autoantibodies and provide specific histone variant-binding reagents.